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Direct interaction of 14-3-3ζ with ezrin promotes cell migration by regulating the formation of membrane ruffle

机译:14-3-3ζ与ezrin的直接相互作用通过调节膜褶的形成促进细胞迁移

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摘要

14-3-3 proteins have been shown to regulate the actin cytoskeleton remodeling, cell adhesion and migration. In this study, we identified ezrin, a cross-linker between plasma membrane and actin cytoskeleton, as a novel 14-3-3ζ interacting partner. The direct interaction between 14-3-3ζ and ezrin was validated in the cells and by in vitro assays. We showed that the 14-3-3ζ binding region in ezrin was located within the N-terminal and central α-helical domains and that the αG-to-αI helices of 14-3-3ζ are responsible for the binding to ezrin. Functional analyses revealed that the regulation of cell migration and membrane ruffling by 14-3-3ζ is ezrin dependent, for which the integrity of ezrin protein was required. Conversely, the knockdown of 14-3-3ζ abrogates also the stimulatory effect of ezrin on cell migration and membrane ruffling. Moreover, we found that the phosphorylation of Thr567 in ezrin facilitates the 14-3-3ζ-ezrin interaction and the formation of membrane ruffles. Taken together, these results suggest strongly that the functions of these two proteins in cell migration are linked and might be mediated by their direct physical interaction, which is important for the formation of membrane ruffles.
机译:已显示14-3-3蛋白可调节肌动蛋白的细胞骨架重塑,细胞粘附和迁移。在这项研究中,我们确定了ezrin(质膜和肌动蛋白细胞骨架之间的交联剂)是一种新型的14-3-3ζ相互作用伴侣。 14-3-3ζ和ezrin之间的直接相互作用在细胞中和通过体外测定得到了验证。我们表明,ezrin中的14-3-3ζ结合区位于N末端和中心α-螺旋结构域内,并且14-3-3ζ的αG-αI螺旋负责与ezrin的结合。功能分析表明,14-3-3ζ对细胞迁移和膜起皱的调节是依斯林依赖性的,为此,需要依斯林蛋白的完整性。相反,敲除14-3-3ζ也会消除ezrin对细胞迁移和膜起皱的刺激作用。此外,我们发现ezrin中Thr567的磷酸化促进了14-3-3ζ-ezrin相互作用和膜褶皱的形成。综上所述,这些结果强烈表明这两种蛋白质在细胞迁移中的功能是相互联系的,并且可能由它们的直接物理相互作用介导,这对形成膜褶很重要。

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